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Bio 10 Enzyme Lab Prep
Prep Instructions Liver Enzyme Juice Schedule: *Start Prep 1-2 weeks before lab *Thaw and pipette 1-2 days before lab Methods: Day 1: For 2 sections, we prepared 50g liver : 150mL DI H2O #Weigh out appropriate amount of liver #Cut into small chunks #Add water and liver chunks to blender #Blend for about 10 minutes in the hood (it stinks) #Pour into TALL SKINNY JARS #Let sit for a half an hour #Freeze until ready to thaw Day 2: #Thaw in refrigerator a day or two before needed #use long pipette and remove the lower, more liquid layer. It can be difficult, maybe spoon some of the top fatty layer off first to avoid contamination. See what works best. #Pipette into tiny flasks. Don't put out more than needed, if left out longer than the say, it will rot and become less effective. Nutrient Agar Plates These plates will be set out for the students to swab during the Enzyme Lab, but they will incubate and be saved for S&C Lab. Prepare 8 plates per section = 1 - control / 1 + control / 6 plates (1 per bench) Prepare them the week before: #Mix agar to the instructions on the agar jar. Agar is located in the media cupboard. #Autoclave #We usually use about 15-18mL agar/plate. SO... 8 plates x # of sections x 18mL per plate = XmL agar total #Maybe split the agar into smaller flasks before pouring plates. DO THIS AS ASEPTICALLY AS POSSIBLE! #Let the agar cool a little but not too much, it solidifies pretty fast. Don't lollygag! #Pour plates near a bunsen burner, leave to set with lids on. #Clearly split each plate into four sections (not controls) and clearly label each plate!!! #Refrigerate in labelled bags until use. #If there is agar left over, pour into a media bottle with an orange top, re-autoclave, label and put in the glass cupboard with the other media. # If you have too many plates, bag them up tightly with tape, label and date the bag and put into the fridge. After Enzyme Lab: # Have the students swab the plates # Incubate the plates 2-3 days # Group and put into Bacterial Fridge in Room 317 until s&C Lab Cotton Swabs These will be used to swab the nutrient agar plates # Take used but not dirty swabs # Put 4 in each "20" test tube: (6 tubes x # of sections = x # of tubes needed (plus a few for good measure) x 4 = # of swabs needed) # autoclave the tubes # Remove and separate for each class section # Cover with cling wrap and refrigerate PLEASE MAKE SURE THE TUBES DO NOT BECOME MOLDY! Buffers You will need pH 2, 4, 7, 10, and 13 buffers in squeeze bottles. Just make sure that there is enough prepared in the reagent cupboard. Check the acid and base cupboards for extra. If one is out, make more per instructions on the capsule bottles. Day-Of Set-up Instructions * Make sure to add calculator to each student station * Put up pH board at buffer station * Only fill student station waters half way * Do not turn on Hot plates * Boiling chips * Put out extra tube rack with clean tubes in the back of the room * Remember cold tubes! H2O with Enzyme on Ice x6 * Eyewear!